Synthetic short DNA probes will be developed for the diagnosis of specific types of Human Papillomavirus (HPV) infections. Short DNA probes of 20-30 nucleotides have the desirable properties of high specificity, fast reaction kinetics, potential high sensitivity and the ability to be synthesized rapidly in large amounts at low cost. Their inherent high specificity is an important attribute. It is possible to design in situ hybridization assays which can detect and discriminate any of the HPV types presently identified. With short DNA probes the contractor will demonstrate that a universal format for in situ hybridization can be developed using 2.4 M Tetraethylammoniuim chloride (TEAC1) in the hybridization or wash steps. This salt solution allows hybridization and hybrid melting reactions to occur solely as a function of probe length. Hence, short probes of the same length (e.g., ca 25 mers) will have the same hybridization and melting properties. In addition, the contractor will demonstrate that agitation of the hybridization and wash steps with ultrasound will significantly decrease the time required for the in situ detection of the various types of HPV from the 24-48 hr now required. The use of short probes in conjunction with TEAC1 salts and ultrasound is expected to be important in the commercialization of DNA probe technology for use in clinical diagnosis.
